Detection of boldenone in the urine of female horses-ex vivo formation versus administration.

Boldenone is an anabolic-androgenic steroid (AAS) that is prohibited in equine sports. However, in certain situations, it is endogenous, potentially formed by the microbes in urine. An approach to the differentiation based on the detection of the biomarkers Δ1-progesterone, 20(S)-hydroxy-Δ1-progesterone and 20(S)-hydroxyprogesterone was assessed, and their concentrations were monitored in the urine of untreated female horses (n = 291) alongside boldenone, boldienone, testosterone and androstenedione. Using an ultra-sensitive analytical method, boldenone (256 ± 236 pg/mL, n = 290) and the biomarkers (Δ1-progesterone up to 57.6 pg/mL, n = 8; 20(S)-hydroxy-Δ1-progesterone 85.3 ± 181 pg/mL, n = 130; 20(S)-hydroxyprogesterone 43.5 ± 92.1 pg/mL, n = 158) were detected at low concentrations. The ex vivo production of Δ1-steroids was artificially induced following the storage of urine samples at room temperature for 7 days in order to assess the concentrations and ratios of the monitored steroids. The administration of inappropriately stored feed source also resulted in an increase in 20(S)-hydroxy-Δ1-progesterone concentrations and the biomarker ratios. Using the results from different datasets, an approach to differentiation was developed. In situations where the presence of boldenone exceeds a proposed action limit of 5 ng/mL, the presence of the biomarkers would be investigated. If Δ1-progesterone is above 50 pg/mL or if 20(S)-hydroxy-Δ1-progesterone is above 100 pg/mL with the ratio of 20(S)-hydroxy-Δ1-progesterone:20(S)-hydroxyprogesterone greater than 5:1, then this would indicate ex vivo transformation or consumption of altered feed rather than steroid administration. There remains a (small) possibility of a false negative result, but the model increases confidence that adverse analytical findings reported in female horses are caused by AAS administrations.

Suppression of reproductive behaviour and gonadal function in female horses-An update.

The behaviour of mares is often detrimental to their performance resulting in frequent demand for methods to suppress gonadal function. In addition, prevention of unintended reproduction especially in feral horse populations may require methods for suppression of gonadal function. Surgical ovariectomy is a safe method but not an acceptable approach in feral mares and undesired in mares where future breeding is considered. There are different approaches for artificial prolongation of the luteal phase resulting in transient inhibition of oestrus and ovulation. Among those, treatment with natural or synthetic progestogens is considered the most common and successful method. Whereas application of intrauterine devices may result in prolongation of luteal function in non-pregnant mares, intrauterine insertion of glass balls is no longer recommended because of complications in individual mares. There are several safer alternatives that may be of interest, especially for population control in free-roaming horses. Treatment with long-acting deslorelin implants inhibited ovulation and oestrus behaviour in mares for limited and variable time intervals in a dose-dependent manner. The effect of GnRH vaccines varies considerably among individual mares, is age dependent, and oestrus-like behaviour may still occur. Contraception via immunization against native porcine or recombinant zona pellucida antigen is successful, but immunocontraception is as much a result of ovarian inactivity as an antibody-based block to sperm-oocyte binding. In conclusion, several treatments for suppression of gonadal function in mares are available, but there are advantages and disadvantages associated that have to be considered. The treatment of choice will thus differ with regard to the demands.